Application of recombinant antibodies for treatment of Clostridioides difficile infection: Current status and future perspective

Abstract

Clostridioides difficile (C. difficile), known as the major cause of antibiotic-associated diarrhea, is regarded as one of the most common healthcare-associated bacterial infections worldwide. Due to the emergence of hypervirulent strains, development of new therapeutic methods for C. difficile infection (CDI) has become crucially important. In this context, antibodies have been introduced as valuable tools in the research and clinical environments, as far as the effectiveness of antibody therapy for CDI was reported in several clinical investigations. Hence, production of high-performance antibodies for treatment of CDI would be precious. Traditional approaches of antibody generation are based on hybridoma technology. Today, application of in vitro technologies for generating recombinant antibodies, like phage display, is considered as an appropriate alternative to hybridoma technology. These techniques can circumvent the limitations of the immune system and they can be exploited for production of antibodies against different types of biomolecules in particular active toxins. Additionally, DNA encoding antibodies is directly accessible in in vitro technologies, which enables the application of antibody engineering in order to increase their sensitivity and specificity. Here, we review the application of antibodies for CDI treatment with an emphasis on recombinant fragment antibodies. Also, this review highlights the current and future prospects of the aforementioned approaches for antibody-mediated therapy of CDI.

Rapid-format recombinant antibody-based methods for the diagnosis of Clostridioides difficile infection: Recent advances and perspectives

“…In contrast to monoclonal antibodies, protein scaffolds are single polypeptide chain structures without disulfide bridges or post-translational modifications that show high solubility and stability and can allow the selection for affinity, stability, and enzymatic activity ( Simeon and Chen, 2018 ; Gebauer and Skerra, 2020 ). Additionally, in vitro antibody display technologies have many advantages, including the ability to be genetically modified to increase selectivity, specificity, and sensitivity, ease of production in E. coli , and reproducibility ( Frenzel et al, 2017 ; Chiu et al, 2019 ; Raeisi et al, 2022a ). Hence, various studies have suggested the use of phage display-derived antibodies for diagnostic purposes ( Chamorro and Merkoçi, 2016 ; Mohd Ali et al, 2021 ).…”

“…Although the hybridoma technology has been successful, it has many shortcomings, e.g., limited number of candidates, time-consuming, inefficiency to generate antibodies toward highly conserved antigens, self-antigens, sensitive antigens, and toxic antigens. A strategy that overcome the disadvantages of hybridoma is techniques based on in vitro antibody production, e.g., yeast display, ribosome display, phage display, and mammalian cell display methods ( Raeisi et al, 2022a ; Valldorf et al, 2022 ), which among them, phage display and ribosome display are preferred in most laboratories due to their large diversity in the range of 10 12 –10 15 variants with high transformation efficiency ( Kunamneni et al, 2020 ; Raeisi et al, 2022a ). Here, we discuss the rAb techniques that are currently being studied for diagnosis proposes.…”

“…The phage display technique has been widely used for detecting various targets, such as haptens, toxins, foreign and self-antigens ( Tullila and Nevanen, 2017 ; Hussack et al, 2018 ; Raeisi et al, 2022a ). For example, a specific antibody against a tumor marker expressed by breast tumor, named ErBb2 protein, has been selected through naïve antibody library that is useful in immunoassay tools ( Bakir et al, 1993 ).…”

“…Additionally, antibiotic therapy for CDI can result in further disruption of the normal gut microbiome and the development of hypervirulent strains of C. difficile . Alternatively, 20–30% of the patients with primary infection symptoms experience recurrent disease within 2–6 weeks after the completion of antibiotic treatment ( Cornely et al, 2012 ; Raeisi et al, 2022a ), which would be higher after secondary and tertiary CDI and more refractory to treatment regimens ( Cornely et al, 2012 ; Eze et al, 2017 ). Therefore, rapid and accurate diagnosis of CDI has been a troublesome issue for disease management, implementation of infection control measures, and epidemiological monitoring ( Burnham and Carroll, 2013 ; Peng et al, 2018 ).…”

“…In recent decades, recombinant antibody (rAb) technologies have widely received much attention as diagnostic tools ( Xu et al, 2017 ; Shali et al, 2018 ; Fouladi et al, 2019 ; Raeisi et al, 2019 ; Alibeiki et al, 2020 ; Raeisi et al, 2020 , 2022b ; Berry et al, 2022 ). The rAb technologies that involve the construction of large libraries of different antibody fragments, such as fragment antigen-binding (Fab), single-chain fragment variable (scFv), minibodies, and nanobodies, can screen antibodies in vitro based on their binding properties, thus help develop more cost-effective antibodies with high sensitivity and specificity ( Bockstaele et al, 2000 ; Angela Chiew Wen et al, 2016 ; Raeisi et al, 2022 ). This review will focus on currently available procedures for CDI diagnosis and different antibody-based rapid detection methods with an emphasis on rAbs.…”